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Literature summary for 3.1.21.10 extracted from
- Functional overlap between the structure-specific nucleases Yen1 and Mus81-Mms4 for DNA-damage repair in S. cerevisiae (2010), DNA Repair, 9, 394-402.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| YEN1 coding sequence amplified from BY4741 genomic DNA and cloned into pDONR221. Further subcloning of the wild-type or mutant into pYES-DEST52 or pAG416GPD-ccdB-HA to generate pYES-DEST52-YEN1-V5-6xHis, pAG416GPD-YEN1-3xHA or pAG416GPDYEN1E193A/E195A-3xHA. Coding sequence of Yen1-V5-6xHis amplified from pYES-DEST52-YEN1-V5-6xHis and subcloned into the BamHI and HindIII sites of p416ADH to generate p416ADH-YEN1-V5-6xHis | Saccharomyces cerevisiae |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| E193A/E195A | catalytically inactive | Saccharomyces cerevisiae |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Saccharomyces cerevisiae | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| DNA + H2O | - |
Saccharomyces cerevisiae | ? | - |
? |  |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Mus81Mms4 | - |
Saccharomyces cerevisiae |
| Yen1 | - |
Saccharomyces cerevisiae |
General Information
| General Information | Comment | Organism |
|---|---|---|
| malfunction | yen1DELTA mutants are repair-proficient. Yen1DELTA mus81ELTA double mutant displays a more severe repair-defective phenotype than the mus81DELTA mutant. Yen1DELTA cells do not exhibit any obvious sensitivity to DNA-damaging agents like the wild-type, whereas yen1DELTA mus81DELTA double mutants are exquisitely sensitive to a variety of DNA-damaging agents that disturb replication fork progression. Yen1DELTA mus81DELTA cells show a hypersensitivity to all agents for which the mus81DELTA single mutant is sensitive: hydroxyurea, 4-nitroquinoline 1-oxide, phleomycin, camptothecin, UV-light, nitrogen mustard and cisplatin. Neither the yen1DELTA or mus81DELTA single mutants nor the yen1DELTA mus81DELTA double mutant show any sensitivity to ionizing radiation up to 200 Gy. Yen1DELTA sgs1DELTA cells are viable and exhibit a similar methyl methanesulfonate, hydroxyurea and 4-nitroquinoline 1-oxide-sensitivity to that observed with the sgs1DELTA single mutant. Toxic recombination intermediates accumulate in the absence of Yen1 and Mus81. After methyl methanesulfonate treatment, yen1DELTA mus81DELTA double mutants arrest with a G2 DNA content and unsegregated chromosomes. Overexpression of Yen1 partially rescues the methyl methanesulfonate sensitivity of mus81DELTA, Yen1 is the only member of the Rad2 family of nucleases that can complement mus81 defects. Yen1ELTA mus81ELTA double mutants are synthetically sick: 15-20 min increase in the duration of the cell cycle of yen1DELTA mus81DELTA double mutants compared with wild-type, yen1DELTA or mus81DELTA cells growing in YPD. Constitutive expression of Yen1-3xHA, but not the catalytically dead version of Yen1, reduces the doubling time of the yen1DELTA mus81DELTA mutant to wild-type levels | Saccharomyces cerevisiae |
| physiological function | Yen1 and Mus81-Mms4 provide alternative and/or overlapping pathways for the repair of methyl methanesulfonate-induced DNA lesions. Yen1 can act upon recombination/repair intermediates that arise in MUS81-defective cells following replication fork damage | Saccharomyces cerevisiae |
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